3. Add 1.5ml/ 35 m Petri dish, allow to set. The plates can be stored at 4°C for up to 1 week.

Top Agar

1.Melt 0.7% Agar (DNA grade agarose) in microwave, cool to 40°C in a water bath. (It is important not to exceed 40°C, otherwise cells will be killed). Also warm 2X DMEM/F12 + additives to the same temperature.

2. Trypsinize cells and count. It is very important to have a positive control line.

5. Label 35mm Petri dishes with base agar appropriately (it is a good idea to remove the plates from 4°C about 30 minutes prior to plating to allow them to warm up to room temperature).

6. For plating add 3ml 2X DMEM/F12 +Additives and 3ml 0.7% Agar to a tube, mix gently and add 1.5ml to each replicate plate (usually plate out in triplicate). NOTE: Only do one tube at a time so that agar does not set prematurely.